Agonizing Hedgehog

Abstract
A new model for the Hedgehog signaling pathway was proposed by Maria Frank-
Kamenetsky and colleagues, and the existence of natural-ligand agonists of the
Hedgehog signaling pathway was hinted at.

Hedgehog history
The Hedgehog family of secreted proteins plays a central role in regulating cell
differentiation and tissue patterning during development. The hedgehog gene (hh)
was first identified in Drosophila embryonic segmentation.
Recently, signaling by Hh has been shown to be important for patterning of the
cerebellum, and misregulated Hh signaling has also been implicated in cancer.
Small-molecule modulators of the Hh pathway might prove more amenable to
pharmacological delivery than the Hh-family proteins themselves.

Sending the signal

Hedgehog signaling is not so simple. The ligand, an extracellular cholesterol-linked
peptide, is processed and cleaved to generate the signaling ligand, which binds to
the cell-surface receptors, activating a transcription factor and inducing a set of
effector genes.
The cellular response to Hh is controlled by two transmembrane proteins, Patched
(Ptc) and Smoothened (Smo), which weave across the cell membrane twelve times
and resemble some transmembrane channels.
Ptc inhibits Smo, and Hh stimulation relieves this inhibition, leading to a nuclear
transcriptional response. Certain mutations in Smo can activate Hh signaling,
bypassing Ptc regulation.

The power of chemical genetics

Frank-Kamenetsky et al.[1] used chemical genetics to identify compounds that could
interfere with the inhibition of Smo by Ptc, or activate Smo independent of Ptc, or act
downstream of Smo.
Frank-Kamenetsky et al. used a high-throughput screen to identify compounds that
block or induce Hh signaling. They found a few candidate agonists that could
stimulate the reporter gene and mimic Hh activity.
Once the cell-based screen was completed, the chemists synthesized 300 derivative
molecules until they found a few compounds that were a thousand times more
effective at eliciting a cellular response.
The cell biologists studied the effects of the agonists on the proliferation of primary
neonatal cerebellar granule neuron precursors and found that they were as effective
as the Hh protein itself.
The researchers found that the agonists activated Hh signaling in Shh-/- embryos,
but had no effect on Hh signaling in embryos lacking Smo. The agonists were
blocked by antagonists that work at the level of Smo or further downstream.
Frank-Kamenetsky et al. found that the agonist had no effect on the stability of the
Ptc protein, but could increase the stability of the Smo receptor.

Learning lessons from drugs

Frank-Kamenetsky et al. have demonstrated that high-throughput chemical genetics
can be used to isolate modulators of a developmental signaling pathway.
Frank-Kamenetsky et al. proposed a new model for Hh signaling based on the
classic 'ternary complex model'. This model predicts that endogenous ligands may
naturally regulate Smo activity.
Cheryll Tickle, from the University of Dundee, UK, studies Hedgehog signaling and
speculates that endogenous molecules act directly on Smoothened, bypassing Ptc.
The study by Frank-Kamenetsky et al.[1] exploits a variety of experimental
techniques to illustrate how high-throughput screening can lead to compound
characterization.